Description:
PepMutePlussiRNA&DNATransfectionReagentisanupgradedversionofPepMutereagent(Cat#SL100566)bycrosslinkingpre-screenedhydrophobicaminoacidstoitsbackbone.WithourproprietarypHDependentConformationalChange(PDCC)technology,thetransfectionreagentwhichcontains36aminoacidslong"speptidebackbonewasformulatedbyadditionofpre-screenedhydrophobicaminoacidstoallaminegroups,conferringPepMutePlusreagentanABIlitytoself-assemblyduringformationoftransfectioncomplexandmakingPepMutereagentaversatileandmostpowerfulgenedeliverytoolwhichprovidesmorethan95%silencingefficiencyat1nMsiRNAinvarietyofmammaliancells.PepMutePlusReagenthavebeenvalidatedtoeffectivelyandreproducIBLytransfectsingleDNAorsiRNAorco-transfectDNA/siRNAtovarietyofmammaliancells.
Figure1.AcartoonshowingPepMutePlussiRNATransfectionReagentwasdeveloped
Size&content:
-PepMutePlusTransfectionReagent,1.0mL,sufficientfor~1000reactionsbasedontransfecting2.5pmolssiRNAin24-wellplate
-PepMutePlusTransfectionBuffer(5x),formulatedformaximaltransfectionefficiency,8.0mL
Storage:
Storeat4°CforPepMutePlusReagentandRTforPepMuteTransfectionBuffer(5x). Ifstoredproperly,theproductisstablefor12monthsorlonger.
Advantages:
-Effectiveknockdown(95%)withonly0.5pmolsiRNA(1.0nMsiRNAin24-wellplate)
-Formulatedforhard-to-transfectmammaliancells
-Onetubereactionandeasytransfectionprotocol
-Bestforbroadspectrumespeciallyhard-to-transfectmammaliancells
-Verylowcytotoxicity
ComparisonsofKnockdownEfficacyofPepMutePlussiRNA&DNATransfectionReagentwithBrandNameProducts
Figure2.KnockdownefficacycomparisonofPepMutePlusTransfectionReagent(upperpanel)vs.Dharmafect4siRNATransfectionReagent(middlepanel)andLipofectamine2000(lowerpanel)onHEK293cells.siRNAtargetingrenillaluciferaseatdifferentfinalconcentrationsrangingfrom0.5to20nMwasco-transfectedwithrenillaluciferasegene(0.5µgofpRL-CMVDNAperwell)bytheabovethreetransfectionreagentspermanufacturers"protocolsintoHEK293cellsgrowingona24-wellplate.Renillaluciferaseactivitywasdetermined24hafterpostco-transfectionwithrenillaluciferasedeterminationsystem(Promega).Theluminescencewasmeasuredfrom5.0µloflysateduring10sintegrationwithaluminometer(BeckmanCoulterLD400).Luciferaseactivitywasexpressedaslightunitsintegratedover10s(RLU)andnormalizedpermgofcellproteinbyusingtheBCAassay.Theerrorsbarsrepresentstandarddeviationderivedfromtriplicateexperiments.Luciferase-silencingefficiencywascalculatedrelativetountreatedcells.WhilePepMutePlusandDharmafect4reagentsdeliveredsignificantgenesilencingfrom1.0nMofrenillaluciferasesiRNA,lipofectamine2000gavegoodknockdownonlyafter30nM(datanotshown).
Figure3.PepMutePlusTransfectionReagentknockeddownGFPgeneexpressioninHepG2cells.GFPCDNA,pEGFP-N3,wasco-transfectwithasiRNAtargetingGFPgene(final5.0nM,rightpanel)andashamsiRNA(final5.0nM,leftpanel)intoHepG2cellsbyPepMutePlusTransfectionReagent.GFPfluorescencewasvisualized24hposttransfectionwithaNikonT2000fluorescencemicorscopy.QuantitativeanalysisshowedthatGFPsiRNAat5.0nMdeliveredbyPepMutePlusTransfectionReagentknockeddown96%co-transfectedGFPexpressioninHepG2cells.
DataSheet&Protocol
-AProtocolforDNA/siRNACo-TranafectiontoMammalianCells
-AProtocolforsiRNATransfectiontoMammalianCells
-AProtocolforDNATransfectiontoMammalianCells
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